Part:BBa_K260010

pMA-BsdR-@CC2FosM

DESCRIPTION: This plasmid backbone is based on pMA from Geneart. It has BioBrick prefix and suffix flanked on both sides by 70 bp of homology arms and a blasticidin resistance cassette (BsdR) for insertion of the contained BioBrick into the proper locus of pCC2FosM ([[Part:BBa_K260000|]]) by recombineering using the HincII fragment including the BioBrick. The product of this reaction is the pCC2FosMB backbone ([[Part:BBa_K260001|]]), which then contains the same BioBrick that was inside the present backbone [[Part:BBa_K260010|]] before, with BioBrick prefix and sufix maintained intact. Initially this was done with the TT_FRT_GFP BioBrick ([[Part:BBa_K260016|]]) using [[Part:BBa_K260010|]]-[[Part:BBa_K260016|]].

The pMA-@CC2FosMB-00500bp backbone is available with [[Part:BBa_K260015|]], which is composed of promoter-FRT-dhfr. If the PvuII fragment from [[Part:BBa_K260004|]]-[[Part:BBa_K260015|]] is transferred by recombineering to the the 00500 locus of [[Part:BBa_K260001|]]-[[Part:BBa_K260016|]], the intervening sequence between the FRT sites of [[Part:BBa_K260015|]] and [[Part:BBa_K260016|]] on the [[Part:BBa_K260001|]] backbone will be 500 bp exactly.

These plasmid backbones are similar and all allow recombineering-mediated transfer of the contained BioBrick (typically [[Part:BBa_K260015|]]) to the pCC2FosMB backbone [[Part:BBa_K2600001|]]. They differ in the homology arms, which determines the locus of insertion into pCC2FosMB. The bp numbers denote final inter-FRT distance as described above:

[[Part:BBa_K260004|]]: pMA-CC2FosMB-00500bp [[Part:BBa_K260005|]]: pMA-CC2FosMB-01000bp [[Part:BBa_K260006|]]: pMA-CC2FosMB-02000bp [[Part:BBa_K260007|]]: pMA-CC2FosMB-05000bp [[Part:BBa_K260008|]]: pMA-RQ-CC2FosMB-10000bp

SOURCE: Geneart plasmid pMA, with 2x 70 bp from pCC2FosMB [[Part:K260001|]].

DESIGN: The 70 bp homologies were designed to delete 100 bp of [[Part:BBa_K260001|]]-[[Part:BBa_K260016|]] and put the FRT site of [[Part:BBa_K260015|]] (if used with the present backbone [[Part:BBa_K260004|]]) at a defined distance from the FRT site of [[Part:BBa_K260016|]].

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal EcoRI site found at 3076
  Illegal NheI site found at 3014
  Illegal SpeI site found at 2
  Illegal PstI site found at 16
  Illegal NotI site found at 9
  Illegal NotI site found at 3082
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal EcoRI site found at 3076
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal prefix found at 3076
  Illegal suffix found at 2
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal prefix found at 3076
  Plasmid lacks a suffix.
  Illegal XbaI site found at 3091
  Illegal SpeI site found at 2
  Illegal PstI site found at 16
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.